Human/Mouse/Rat DOK7 Antibody

Detection of Human, Mouse, and Rat DOK7 by Western Blot. Western blot shows lysates of untreated MCF-7 human breast cancer cell line and T47D human breast cancer cell line and C2C12 mouse myoblast cell line and L6 rat myoblast cell line untreated (-) or treated (+) with 2% horse serum for 6 days. PVDF Membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat DOK7 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6398) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for DOK7 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Detection of Human DOK7 by Simple Western^TM. Simple Western lane view shows lysates of MCF-7 human breast cancer cell line and T47D human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for DOK7 at approximately 60 & 62 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat DOK7 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6398) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Mouse DOK7 by Western Blot Sorbs1 is enriched at AChR aggregates, and Sorbs1 RNAi blocks AChR clustering in vitro. (A) Treatment of myotubes with siRNA directed against Sorbs1 blocks AChR clustering in C2C12 cells. Montages containing sixteen fields at a magnification of 10× were analyzed with ImageJ software (NIH). (B) Sorbs1 siRNA significantly reduces Sorbs1 protein expression in myotubes. (C) Sorbs1 protein is highly enriched at sites where AChRs aggregate. (D) Agrin stimulates tyrosine phosphorylation of MuSK and Dok-7 at similar levels in myotubes treated with Sorbs1 siRNA. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26527617), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of DOK7 by Western Blot hDOK7 overexpression stimulates long-term elevation of DOK7 protein expression in mouse hindlimb muscles but not the heart(A) Western blots of tissue lysates probed with anti-DOK7 antibody shown at 55 kDa (middle band). Tissue from three mice each untreated or injected with high- or low-dose AAV9-tMCK-DOK7 was run alongside internal standards (ISs) produced in triplicate, enabling cross-gel comparison. Tissues were taken from mice at 3 months (M) and 6 months of age.(B–D) Bar charts comparing the effect of dose on tissue-specific DOK7 expression. Western blot data were quantified and normalized to the average value of three internal standards. DOK7 expression was significantly increased at 3 and 6 months following high-dose treatment in the hindlimb soleus muscle (B) and lumbricals (C) but not in heart tissue (D), when compared to untreated controls. Specifically, there was a 3.8-fold increase in DOK7 expression in the soleus 3 months following treatment and a 1.9-fold increase at 6 months (B; p < 0.0001 and p = 0.019), while in the lumbricals, there was a 4.7-fold increase in DOK7 expression at 3 months and a 4.6-fold increase at 6 months (C; p = 0.0009 and p = 0.0031) (one-way ANOVAs with Šídák’s multiple comparisons, means of n = 3 per group [with SEM], and each data point represents one animal). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37637210), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of DOK7 by Western Blot AAV9-tMCK-DOK7 increases DOK7 expression in skeletal muscle(A) Schematic of the AAV9-tMCK-DOK7 plasmid showing the skeletal muscle targeting tMCK promoter & DOK7 transgene.(B–D) DOK7 expression across a range of muscles & organs from 1-month-old mice treated with high dose (5e11 vg), low dose (1.25e11 vg), or untreated with AAV9-tMCK-DOK7. “IS” denotes internal standards run in triplicate used to allow comparison between blots. (B) Western blots where the molecular weight of DOK7 is indicated. Dotted lines illustrate where blot sections have been cropped together. (C) Bar graph of endogenous expression in skeletal muscle & tissue shown by quantified WBs. Values are normalized to the average of three DOK7 internal standards (ISs). Each dot represents a single mouse, & bars represent the mean (with SEM) (n = 3 per bar). (D) DOK7 expression in response to high-dose or low-dose treatments were compared by one-way ANOVA with Holm-Šídák’s multiple comparisons. Each bar represents an average of three animals (with SEM), normalized to internal standard (IS) & then normalized to endogenous protein levels for that tissue. Expression increased by 40x over endogenous levels in the GC (p = 0.0114), & the TA (p = 0.0242), hindlimb lumbricals (p = 0.0074), & TVA (p = 0.013) each showed a marked rise in DOK7. The soleus, which exhibited the highest endogenous levels of DOK7 in skeletal muscle, still expressed ∼4x as much DOK7 following high-dose treatment (p = 0.0032). There was also a small but significant increase in DOK7 expression in the heart following high-dose AAV9-tMCK-DOK7 treatment (p = 0.0018, n = 3 in all groups, one-way ANOVAs with Holm-Šídák’s comparisons). SPC, spinal cord; TVA, transverse abdominis; TA, tibialis anterior; GC, gastrocnemius; ∗p < 0.05, ∗∗p < 0.01. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37637210), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of DOK7 by Western Blot AAV9-tMCK-DOK7 increases DOK7 expression in skeletal muscle(A) Schematic of the AAV9-tMCK-DOK7 plasmid showing the skeletal muscle targeting tMCK promoter & DOK7 transgene.(B–D) DOK7 expression across a range of muscles & organs from 1-month-old mice treated with high dose (5e11 vg), low dose (1.25e11 vg), or untreated with AAV9-tMCK-DOK7. “IS” denotes internal standards run in triplicate used to allow comparison between blots. (B) Western blots where the molecular weight of DOK7 is indicated. Dotted lines illustrate where blot sections have been cropped together. (C) Bar graph of endogenous expression in skeletal muscle & tissue shown by quantified WBs. Values are normalized to the average of three DOK7 internal standards (ISs). Each dot represents a single mouse, & bars represent the mean (with SEM) (n = 3 per bar). (D) DOK7 expression in response to high-dose or low-dose treatments were compared by one-way ANOVA with Holm-Šídák’s multiple comparisons. Each bar represents an average of three animals (with SEM), normalized to internal standard (IS) & then normalized to endogenous protein levels for that tissue. Expression increased by 40x over endogenous levels in the GC (p = 0.0114), & the TA (p = 0.0242), hindlimb lumbricals (p = 0.0074), & TVA (p = 0.013) each showed a marked rise in DOK7. The soleus, which exhibited the highest endogenous levels of DOK7 in skeletal muscle, still expressed ∼4x as much DOK7 following high-dose treatment (p = 0.0032). There was also a small but significant increase in DOK7 expression in the heart following high-dose AAV9-tMCK-DOK7 treatment (p = 0.0018, n = 3 in all groups, one-way ANOVAs with Holm-Šídák’s comparisons). SPC, spinal cord; TVA, transverse abdominis; TA, tibialis anterior; GC, gastrocnemius; ∗p < 0.05, ∗∗p < 0.01. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37637210), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of DOK7 by Western Blot hDOK7 overexpression stimulates long-term elevation of DOK7 protein expression in mouse hindlimb muscles but not the heart(A) Western blots of tissue lysates probed with anti-DOK7 antibody shown at 55 kDa (middle band). Tissue from three mice each untreated or injected with high- or low-dose AAV9-tMCK-DOK7 was run alongside internal standards (ISs) produced in triplicate, enabling cross-gel comparison. Tissues were taken from mice at 3 months (M) and 6 months of age.(B–D) Bar charts comparing the effect of dose on tissue-specific DOK7 expression. Western blot data were quantified and normalized to the average value of three internal standards. DOK7 expression was significantly increased at 3 and 6 months following high-dose treatment in the hindlimb soleus muscle (B) and lumbricals (C) but not in heart tissue (D), when compared to untreated controls. Specifically, there was a 3.8-fold increase in DOK7 expression in the soleus 3 months following treatment and a 1.9-fold increase at 6 months (B; p < 0.0001 and p = 0.019), while in the lumbricals, there was a 4.7-fold increase in DOK7 expression at 3 months and a 4.6-fold increase at 6 months (C; p = 0.0009 and p = 0.0031) (one-way ANOVAs with Šídák’s multiple comparisons, means of n = 3 per group [with SEM], and each data point represents one animal). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37637210), licensed under a CC-BY license. Not internally tested by R&D Systems.